Biological basis for restriction of microRNA targets to the 3′ untranslated region in mammalian mRNAs
Top Cited Papers
Open Access
- 1 February 2009
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Structural & Molecular Biology
- Vol. 16 (2), 144-150
- https://doi.org/10.1038/nsmb.1552
Abstract
MicroRNAs (miRNAs) interact with target sites located in 3' untranslated regions (3'UTR) of mRNAs to down-regulate their expression when the appropriate miRNA is bound to target mRNA. To establish the functional importance of target localization in the 3' UTR, we modified the stop codon to extend the coding region of the transgene reporter through the miRNA target sequence. As a result, the miRNAs lost their ability to inhibit translation but retained their ability to function as siRNAs in mammalian cells in culture and in vivo. The addition of rare but not optimal codons upstream of the extended opening reading frame (ORF) made the miRNA target more accessible and restored miRNA-induced translational knockdown. Taken together, these results suggest that active translation impedes miRNA-programmed RISC association with target mRNAs, and support a mechanistic explanation for the localization of most miRNA target sites in noncoding regions of mRNAs in mammals.Keywords
This publication has 49 references indexed in Scilit:
- siRNAs can function as miRNAsGenes & Development, 2003
- A microRNA in a Multiple-Turnover RNAi Enzyme ComplexScience, 2002
- Both Natural and Designed Micro RNAs Can Inhibit the Expression of Cognate mRNAs When Expressed in Human CellsMolecular Cell, 2002
- Regulation of c-myc mRNA Decay by Translational Pausing in a Coding Region Instability DeterminantMolecular and Cellular Biology, 2002
- Identification of Tissue-Specific MicroRNAs from MouseCurrent Biology, 2002
- Micro RNAs are complementary to 3′ UTR sequence motifs that mediate negative post-transcriptional regulationNature Genetics, 2002
- Two Genetic Circuits Repress the Caenorhabditis elegans Heterochronic Gene lin-28 after Translation InitiationDevelopmental Biology, 2002
- Detection of Antisense and Ribozyme Accessible Sites on Native mRNAs: Application to NCOA3 mRNAMolecular Therapy, 2001
- Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon systemNature Genetics, 2000
- The lin-4 Regulatory RNA Controls Developmental Timing in Caenorhabditis elegans by Blocking LIN-14 Protein Synthesis after the Initiation of TranslationDevelopmental Biology, 1999